毛冬青甲素对大鼠脑缺血再灌注后VEGF和VEGFR2的表达及神经元再生的影响
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摘要
目的:探讨毛冬青甲素(Ilexonin,IA)对大鼠脑缺血再灌注后VEGF和VEGFR2的表达及神经元再生的影响的可能作用机制。
方法:线栓左侧大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)制成SD大鼠脑缺血模型,随机分成:正常组、假手术组、模型组、IA治疗组(分IA71、142、285μmol·kg- 1三个亚组)等4组。采用Longa评分法进行大鼠神经功能评分; TTC染色法观察脑缺血再灌注模型组和治疗组缺血灶情况;免疫组化检测缺血再灌注后不同时段各组缺血周边区VEGF、VEGFR2阳性细胞数的变化;免疫荧光检测缺血再灌注后不同时段各组脑侧室室管膜下区(SVZ)神经元再生标记物Nestin阳性细胞数的变化;免疫印迹法检测缺血再灌注后不同时段各组缺血周边区VEGF、VEGFR2及脑侧室室管膜下区(SVZ)神经元再生标记物Nestin蛋白的表达情况。并比较不同剂量(IA71、142、285μmol·kg- 1)的IA干预对大鼠神经功能缺损评分,缺血区周边组织VEGF、VEGFR2及SVZ的Nestin蛋白表达和阳性细胞数的影响。
结果:脑缺血再灌注后3d,模型大鼠神经功能缺损评分较高(P<0.05 VS 14d、28d);VEGF、VEGFR2、Nestin阳性细胞在正常组及假手术组散在见到,模型组7d达到高峰,随时间延长其表达逐渐减少;VEGF、VEGFR2、Nestin蛋白于正常组及假手术组表达较少,模型组自第3d有一定量的表达,第7天表达最多,之后逐渐减少,至28天仍有表达。模型组7d与相应模型组其它时段比较P<0.05。经不同剂量IA71、142、285μmol·kg- 1干预后,大鼠神经功能缺损评分有明显下降;缺血周边区VEGF、VEGFR2及SVZNestin蛋白表达和阳性细胞数明显增多,与模型组相应时间点比较显著性差异(P<0.05)。以285μmol·kg- 1作用为著。
结论:1、脑缺血再灌注损伤可诱导VEGF、VEGFR2的早期表达(至少14d), VEGF、VEGFR2的早期应激表达可能与及时挽救缺血周边组织神经元的存活和再生进而促进神经功能恢复有关。2、毛冬青甲素可以促进缺血周边组织VEGF、VEGFR2表达(至少14d),其机制可能通过上调VEGF、VEGFR2表达促进神经元的存活和再生进而发挥其神经保护作用。
OBJECTIVE To explore the effect of IlexoninA on expression of VEGF, VEGFR2 and neurogenesis and its possible mechanism after cerebral ischemia- reperfusion in Rats.
METHODS The model of Middle cerebral artery occlusion(MCAO) was established to transient focal ischemia by placement of an intraluminal filament at the origin of left middle cerebral artery. The rats were divided randomly into: control, sham operation, model and IlexoninA (IA71、142、285μmol·kg- 1) treatment groups. Neuronogical behavior was evaluated by Longa’s scoring method. TTC staining was applied to observe cerebral ischemia in MCAO model group and IlexoninA treatment groups. Immunohistochemistry were used to observe the expression of VEGF, VEGFR2 positive cells in the ischemic brain tissue in theat different time of ischemia and reperfusion. Immunofluorescence were used to observe the expression of Nestin (neurogenesis Marker)positive cells in the SVZ in theat different time of ischemia and reperfusion.Western bloting were used to observe the expression of VEGF, VEGFR2 protein in the ischemic brain tissue and Nestin protein(neurogenesis Marker) in the subventricular zone in theat different time of ischemia and reperfusion.The effects of IA intervention (71、142、285μmol·kg- 1) on neurological deficit score and the expression of VEGF, VEGFR2 protein in ischemia surrounding areas and Nestin protein in ischemia SVZ were mainly quantified.And the effects of IA intervention (71、142、285μmol·kg- 1) on the positive cells of VEGF, VEGFR2 in ischemia surrounding areas and Nestin in ischemia SVZ were mainly quantified.
RESULTS Neurological deficits of model were more at 3d(P<0.05 compared to 14d and 28d).In control and sham groups, there were scattered VEGF, VEGFR2 positive cells (mainly in the cortex), and the number of VEGF, VEGFR2 positive cells were peaked at 7d in model group. With the prolongation of reperfusion, the expression of VEGF, VEGFR2 decreased.VEGF, VEGFR2 protein in the control group and sham group was much less than model group.In model group, there was a certain amount at 3d after reperfusion, reached to peak at 7d, then reduced gradually, to 28d was still expression. Similarly, Nestin protein reached its peak at 7d after reperfusion, and then decrcased gradually, compared to other model times (P<0.05). After the intervention of IA71、142、285μmol·kg- 1, the neurological deficit score decreased significantly, and the expression of VEGF, VEGFR2 protein in ischemia surrounding areas and Nestin in ischemia SVZ were increased comparing to model group at corresponding time points (P<0.05), and the positive cells of VEGF, VEGFR2 in ischemia surrounding areas and Nestin in ischemia SVZ were increased comparing to model group at corresponding time points (P<0.05), with pronounced effct at dose of IA285μmol·kg- 1.
CONCLUSION
1.Cerebral ischemia reperfusion injury induced the expression of VEGF, VEGFR2 in the early time(at least 14d). It is possible that the early expression of VEGF, VEGFR2 were involved to promote the survival and remodeling of peripheral neurons ,with promoting the recovery of neurological function.
2. IlexoninA can promote the the expression of VEGF, VEGFR2 in the early time(at least 14d).It may be passed through upregulation of VEGF, VEGFR2 to promote the repair and regeneration of neurons to play its role of neuroprotective.
方法:线栓左侧大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)制成SD大鼠脑缺血模型,随机分成:正常组、假手术组、模型组、IA治疗组(分IA71、142、285μmol·kg- 1三个亚组)等4组。采用Longa评分法进行大鼠神经功能评分; TTC染色法观察脑缺血再灌注模型组和治疗组缺血灶情况;免疫组化检测缺血再灌注后不同时段各组缺血周边区VEGF、VEGFR2阳性细胞数的变化;免疫荧光检测缺血再灌注后不同时段各组脑侧室室管膜下区(SVZ)神经元再生标记物Nestin阳性细胞数的变化;免疫印迹法检测缺血再灌注后不同时段各组缺血周边区VEGF、VEGFR2及脑侧室室管膜下区(SVZ)神经元再生标记物Nestin蛋白的表达情况。并比较不同剂量(IA71、142、285μmol·kg- 1)的IA干预对大鼠神经功能缺损评分,缺血区周边组织VEGF、VEGFR2及SVZ的Nestin蛋白表达和阳性细胞数的影响。
结果:脑缺血再灌注后3d,模型大鼠神经功能缺损评分较高(P<0.05 VS 14d、28d);VEGF、VEGFR2、Nestin阳性细胞在正常组及假手术组散在见到,模型组7d达到高峰,随时间延长其表达逐渐减少;VEGF、VEGFR2、Nestin蛋白于正常组及假手术组表达较少,模型组自第3d有一定量的表达,第7天表达最多,之后逐渐减少,至28天仍有表达。模型组7d与相应模型组其它时段比较P<0.05。经不同剂量IA71、142、285μmol·kg- 1干预后,大鼠神经功能缺损评分有明显下降;缺血周边区VEGF、VEGFR2及SVZNestin蛋白表达和阳性细胞数明显增多,与模型组相应时间点比较显著性差异(P<0.05)。以285μmol·kg- 1作用为著。
结论:1、脑缺血再灌注损伤可诱导VEGF、VEGFR2的早期表达(至少14d), VEGF、VEGFR2的早期应激表达可能与及时挽救缺血周边组织神经元的存活和再生进而促进神经功能恢复有关。2、毛冬青甲素可以促进缺血周边组织VEGF、VEGFR2表达(至少14d),其机制可能通过上调VEGF、VEGFR2表达促进神经元的存活和再生进而发挥其神经保护作用。
OBJECTIVE To explore the effect of IlexoninA on expression of VEGF, VEGFR2 and neurogenesis and its possible mechanism after cerebral ischemia- reperfusion in Rats.
METHODS The model of Middle cerebral artery occlusion(MCAO) was established to transient focal ischemia by placement of an intraluminal filament at the origin of left middle cerebral artery. The rats were divided randomly into: control, sham operation, model and IlexoninA (IA71、142、285μmol·kg- 1) treatment groups. Neuronogical behavior was evaluated by Longa’s scoring method. TTC staining was applied to observe cerebral ischemia in MCAO model group and IlexoninA treatment groups. Immunohistochemistry were used to observe the expression of VEGF, VEGFR2 positive cells in the ischemic brain tissue in theat different time of ischemia and reperfusion. Immunofluorescence were used to observe the expression of Nestin (neurogenesis Marker)positive cells in the SVZ in theat different time of ischemia and reperfusion.Western bloting were used to observe the expression of VEGF, VEGFR2 protein in the ischemic brain tissue and Nestin protein(neurogenesis Marker) in the subventricular zone in theat different time of ischemia and reperfusion.The effects of IA intervention (71、142、285μmol·kg- 1) on neurological deficit score and the expression of VEGF, VEGFR2 protein in ischemia surrounding areas and Nestin protein in ischemia SVZ were mainly quantified.And the effects of IA intervention (71、142、285μmol·kg- 1) on the positive cells of VEGF, VEGFR2 in ischemia surrounding areas and Nestin in ischemia SVZ were mainly quantified.
RESULTS Neurological deficits of model were more at 3d(P<0.05 compared to 14d and 28d).In control and sham groups, there were scattered VEGF, VEGFR2 positive cells (mainly in the cortex), and the number of VEGF, VEGFR2 positive cells were peaked at 7d in model group. With the prolongation of reperfusion, the expression of VEGF, VEGFR2 decreased.VEGF, VEGFR2 protein in the control group and sham group was much less than model group.In model group, there was a certain amount at 3d after reperfusion, reached to peak at 7d, then reduced gradually, to 28d was still expression. Similarly, Nestin protein reached its peak at 7d after reperfusion, and then decrcased gradually, compared to other model times (P<0.05). After the intervention of IA71、142、285μmol·kg- 1, the neurological deficit score decreased significantly, and the expression of VEGF, VEGFR2 protein in ischemia surrounding areas and Nestin in ischemia SVZ were increased comparing to model group at corresponding time points (P<0.05), and the positive cells of VEGF, VEGFR2 in ischemia surrounding areas and Nestin in ischemia SVZ were increased comparing to model group at corresponding time points (P<0.05), with pronounced effct at dose of IA285μmol·kg- 1.
CONCLUSION
1.Cerebral ischemia reperfusion injury induced the expression of VEGF, VEGFR2 in the early time(at least 14d). It is possible that the early expression of VEGF, VEGFR2 were involved to promote the survival and remodeling of peripheral neurons ,with promoting the recovery of neurological function.
2. IlexoninA can promote the the expression of VEGF, VEGFR2 in the early time(at least 14d).It may be passed through upregulation of VEGF, VEGFR2 to promote the repair and regeneration of neurons to play its role of neuroprotective.
引文
[1]Lendahl U,Zimmerman LB,McKay RD.CNS stem cells express a new class of intermediate filament protein[J].Cell,1990,6O(4):585—595.
[2] Doetsch F.A niche for adult neural stem cells[J].Curr Opin Genet Dev,2003,13(5):543—550.
[3] Meister B,Bautz F,Brugger W,et a1.Expression of vascular endothelial growth factor(VEGF)and its receptors in human Neuroblastoma[J].Eur J Cancer,1999,35(3):445—449.
[4]李欣,张君,梁尚栋.血管内皮生长因子在神经系统中的作用与机制[J].神经解剖学杂志,2010, 26(5): 561—563.
[5]华海婴,李艳瑛.毛冬青甲素药理作用的研究[J].中国现代医药杂志,2006,8(11) :137—138.
[6]罗新海.毛冬青甲素治疗缺血性脑血管病. [J].广东医学, 1985, 64:26—28.
[7]江苏新医学院编.中医药大辞典.上册.[S]上海人民出版社, 1977: 441—443.
[8] Nacher J,Rosell DR,Alonso-Liosa G,MeEwen BS. NMDA receptor antagonist treatment induces a long-lasting increase in the number of proliferating cells PSA-NCAM-immunoreactive granule neurons and radial glia in the adult rat dentate gyrus.Eur J Neurosci.2001,13(3):512—520.
[9]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without cranicetomy in rats[J].Stroke,1989,20(1):84—91.
[10]方舒东,朱也森.脑缺血再灌注损伤的病理生理研究进展[J].医学综述,2006,12(18):1114—1116.
[11]韩敏,刘艳凯,薛茜.脑缺血再灌注损伤的研究进展[J].河北北方学院学报,2006,23(5):75—78.
[12]冯加纯,饶明俐,张淑琴,等.大鼠全脑缺血再灌注后几个脑区在不同时间的LPO、SOD、GSH-Px含量变化[J].中风与神经疾病杂志,1994,11(3):129—132.
[13]郭玉璞.缺血性脑损伤的病理生理基础和脑梗塞的治疗原则[J].基础医学与临床,1998,18(2):1—4.
[14]Lledo PM ,Alonso M , Grubb MS, et al . Adult neurogenesis and functional plasticity in neuronal circuits . Nat Rev Neurosci, 2006,7: 179—193.
[15]Alvarez-Buylla A, Garcia-Verdugo JM. Neurogenesis in adult subventricular zone. J Neurosci. 2002 ,22(3):629-634.
[16]Pencea V, Bingaman KD, Freedman LJ, Luskin MB. Neurogenesis in the subventricular zone and rostral migratory stream of the neonatal and adult primate forebrain. Exp Neurol. 2001,172(1):1-16.
[17]Kornack DR, Rakic P. The generation, migration, and differentiation of olfactory neurons in the adult primate brain.Proc Natl Acad Sci U S A. 2001,98(8):4752-4757.
[18]Sehara Y, Hayashi T, Deguchi K, et al . Distribution of inducible nitric oxide synthase and cell proliferation in rat brain after transient middle cerebral artery occlusion. Brain Res ,2006 , 1093 : 190—197.
[19]Li Y,Chopp M.Temporal profile of nestin expression after focal cerebral ischemia in adult rat[J].Brain Res,1999,838(1-2):1—10.
[20]Barres BA. New roles for glia[J]. J Neurosci, 1991, 11 (12) :3685—94.
[21]王娟,周义成,陈信坚.脑静脉窦血栓形成的MRI诊断[J].中国临床医学影像学杂志,2005,16(1):1008—1062.
[22] Loubeyre P,De Jaegere T,Tran-Minh VA.Three-dimensional phase contrast MR cerebral venography with zero filling interpolation in the slice encoding direction[J].Magn Reson Imaging,1999,17(8):1227—1233.
[23]Chiasson BJ,Tropepe V,Morshead CM ,van der Kooy D.Adult mammalian forebrain ependymal and subependymal cells demonstrate proliferative potentia1,but only subependymal cells have neura1 stem cell characteristics[J].J Neurosci,1999,19(11):4462—4471.
[24]Stevenson J,Knopp EA,Litt AW .MP-RAGE subtraction venography:a new technique[J].J Magn Reson Imaging,1995,5(2):239—241.
[25] Doetsch F.A niche for adult neural stem cells[J].Curr Opin Genet Dev,2003,13(5):543—550.
[26] Meister B,Bautz F,Brugger W,et a1.Expression of vascular endothelial growth factor(VEGF)and its receptors in human Neuroblastoma[J].Eur J Cancer,1999,35(3):445—449.
[27]焦淑洁,许慧芳,许杰,等. VEGF在人胚胎干细胞向神经元分化中作用的研究[J].Stroke and Nervous Diseases,2009,16(3):134—137.
[28]Hansen TM, Moss AJ, Brindle NP. Vascular endothelial growth factor and angiopoietins in neurovascular regeneration and protection following stroke[J]. Curr Neurovasc Res, 2008, 5(4): 236—245.
[29]Shen F, Fan Y, Su H, et al. Adeno-associated viral vector-mediated hypoxia-regulated VEGF gene transfer promotes angiogenesis following focal cerebral ischemia in mice[J]. Gene Ther, 2008,15(1): 30—39.
[30] Svensson B,Peters M,Poppe M,et a1.Vascular endothelial growth factor protects cultured rat hippocampal neurons against hypoxic injury via an antiexcitotoxic,caspase-independent mechanism[J].J Cereb Blood Flow Metab,2002,22(10):1170—1175.
[31]Sun Y,Jin K,Xie L,et a1.VEGF-induced neuroprotection,neurogenesis,and angiogenesis after focal cerebral ischemia[J].J Clin Invest,2003 Jun,111(12):1843—1851.
[32]杨树法等,血管内皮生长因子对胚胎干细胞来源神经干细胞增殖的促进[J].中国临床康复2006,10(1);12—14.
[33]彭好等,神经生长因子对新生鼠缺氧缺血性脑损伤神经修复作用的研究[J].中国新生儿科杂志,2007,22(2):85—88.
[34]董斌,罗其中,徐英辉.脑缺血半暗带区研究进展[J].中华神经外科疾病研究杂志,2004,4(1):90—92.
[35] Higashida RT,Furlan AJ,Roberts H,et al.Trial design and reporting standards for intra-arterial cerebral thrombolysis for acute ischemic stroke[J].Stroke,2003,34(8):109—137.
[36]陈洁文,谭宝璇,苏文.毛冬青甲素对脑细胞缺氧损伤的保护及其机制的实验研究[J].广州中医药大学学报,1997,14(2):97.
[37] Magavi SS, Macklis JD.Manipulation of neural precursorsin situ tower induction of neurogenesis in the adult brain:potential and limitation and limitation.Clin Neurosci Res,2002, 2 (1-2) : 40—57.
[38] Zhang PB, L iu Y, L i J , et al. Ependymal / subventricular zone cells migrate to the peri-infarct region and differentiate into neurons and astrocytes after focal cerebral ischemia in adult rats. Di Yi Jun YiDa Xue Xue Bao, 2005,25 (10) : 1201—1206.
[39] Yan JP, Liu XF, Liu H J, et al. Extracellular signal regulated k-inase involved in NGF /VEGF induced neuroprotective effect[J].Neurosci Lett, 2008, 434: 212—217.
[1] Tsyb AF, Roshal LM, Yuzhakov VV, et al. Morphofunctional study of the therapeutic effect of autologous mesenchymal stem cells in experimental diffuse brain injury in rats.Bull Exp Biol Med 2006;142(1):140—147.
[2] Wacharaprechanont T. Fetal stem cell: from research to clinical use.J Med Assoc Thai 2005;88 Suppl 2:S133—137.
[3]zhang YW, Denham J,Thies RS. Oligodendrocyte progenitor cells derived from human embryonic stem cells express neurotrophic factors.Stem Cells Dev 2006;15(6): 943—952.
[4] Li Y, Chopp M, Chen J et al. Intrastriatal transplantation of bone marrow nonhematopoietic cells improves functional recovery after stroke in adult mice. J Cereb Blood Flow Metab 2000;20(9):1311—1319.
[5] Sanchez-Ramos JR, Song S, Kamath SG,et al. Expression of neural markers in human umbilical cord blood. Exp Neurol 2001; 171(1):109—115.
[6] Chen J, Sanberg PR, Li Y, et al. Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats.Stroke 2001;32: 2682—2688.
[7]Yang L, Zheng J, Wang C. The in vitro study of the human adipose tissue-derived stromal cells differentiating into the neuron-like cells.Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2006;20(8):783—786.
[8] Rehman J, Traktuev D, Li J,ET al. Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation2004;109:1292—1298.
[9] Lathia JD,Rao MS,Mattson MP,et al. The microenvironment of the embryonic neural stem cell: Lessons from adult niches?Dev Dyn 2007;236(12):3267—3282.
[10] Hwang WS, Roh SI, Lee BC et al. Patient-specific embryonic stem cells derived from human SCNT blastocysts. Science 2005;308:1777—1783.
[11] Priddle H, Jones DK, Burridge PW, et al. Hematopoiesis from human embryonic stem cells: overcoming the immune barrier in stem cell therapies. Stem cells 2006;24(4):815—824.
[12] Bieberich E, Silva J, Wang G, et al. Selective apoptosis of pluripotent mouse andhuman stem cells by novel ceramide analogues prevents teratoma formation and enriches for neural precursors in ES cell-derived neural transplants.J Cell Biol 2004;167:723—734.
[13] Coyne TM, Marcus AJ, Woodbury D,et al. Marrow stromal cells transplanted to the adult brain are rejected by an inflammatory response and transfer donor labels to host neurons and glia. Stem Cells 2006;24(11):2483—2492.
[14] Ruth Kirschstein MD. Assessing Human Stem Cell Safety stem cells:scientific progress and future research ditections. Washington,DC, USA: Department of Health and Human Services 2001:93—99. [15 ]Chen J , Li Y, Chopp M. Intracerebral transplantation of bone marrow with BDNF after MCAO in rat. Neuropharmacology 2000; 39:711—716.
[16] Zhao LR, Duan WM , Reyes M , et al. Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp Neurol 2002;174(1):11—20. [17 ]Savitz SI, Rosenbaum DM , Dinsmore JH, et al. Cell transplantation for stroke . Ann Neurol 2002;52: 266—275.
[18] Chen J , Li Y, Wang L, et al. Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke 2001; 32(4): 1005—1011.
[19] Newman MB, Willing AE, Manresa JJ, et al. Cytokines produced by cultured human umbilical cord blood (HUCB) cells: implications for brain repair Exp Neurol 2006 ;199(1):201—208.
[20] Kondziolka D, Wechsler L, Goldstein S, et al. Transplantation of cultured human neuronal cell for patients with stroke.Neurology 2000;55: 505—569.
[21] Wiiling AE , Vendrme M , Mallery J ,et al. Mobilized peripheral blood cells administered intravenously produce functional recovery in stroke.Cell Transplant 2003;12:449—454.
[22] Lee ST, Park JE , Lee K, et al. Noninvasive method of immortalized neural stem-like cell transplantation in an experimental model of Huntington,s disease. J Neurosci Methods 2006;152(1-2):250—254.
[23] Jin K, Sun Y, Xie L , et al. Comparison of ischemia directed migration of neural precursor cells after intrastriatal , intraventricular , or intravenous transplantation in the rat. Neurobiol Dis 2005;18:366—374.
[24]Verfaillie CM,SchwartI R,Reyes M,et al.Unexpected potential of adult stell cell[J].Ann NY Acad Sci,2003,996:231—234.
[25] Shyu WC,Lin SZ,Ching MF,et al.Intracerbral peripheral blood stem cell(CD34+)implantation induces Neuroplasticity by enhancing betal 1 integrin-mediated angiogenesis in chronic stroke rats[J].Neurosci, 2006.26(13): 3444—3453. [26 ]Li Y.Chen J,Zhang CL,et al.Gliosis and brain remodeling after treatment of stroke in rats with marrow stromal cells. Glia.2005,49(3):407—417.
[27] Zhu D Y,Lau L,Liu SH,et al.Activation of camp-response-element–binding protein(CREB)after focal cerebral ischemia stimulates neurogenesis in the adult dentate gyrus.Proc Natl Acad Sci USA,2004,101:9453—9457.
[28] Xin H,Li Y,Chen X,et al.Bone marrow stromal cells induce BMP2/4 production in oxygen-glucose-deprived astrocytes,which promotes an astrocytic phenotype in adult subventricular progenitor cells[J].Neurosci Res,2006,83(8):1485—1493.
[29] Zhang C,Li Y,Chen J,et al.Bone marrow stromal cells upregulate expression of bone morphogenetic proteins 2 and 4 gap junction protein connexin-43 and synaptophysin after stroke in rats[J].Neuroscience,2006,141(2):687—695.
[30] Hess DC,Abe T,Hill WD,et al.Hematopoietic origin of microglial and perivascular cells in brain[J],Exp Neural, 2004;186(2):134—144.
[31] Chen J,Li Y,Katakowski M,et al.Intravenous bone marrow stromal cell therapy reduces apoptosis and promotes endogenous cell proliferation after stroke in female rat[J].Neurosci Res,2003,73:778—786.
[32]步星耀,黄志起,张永福.骨髓基质细胞的生物学特性及应用研究[J].实用诊断与治疗杂志,2004,18(1):35—38.
[33] Bang OY,Lee,IS,Lee PH,et a1.Autologous mesenchymal stem cell transp1antati。n in stroke patients [J]. Ann Neurol,2005,57(6):874—882.
[2] Doetsch F.A niche for adult neural stem cells[J].Curr Opin Genet Dev,2003,13(5):543—550.
[3] Meister B,Bautz F,Brugger W,et a1.Expression of vascular endothelial growth factor(VEGF)and its receptors in human Neuroblastoma[J].Eur J Cancer,1999,35(3):445—449.
[4]李欣,张君,梁尚栋.血管内皮生长因子在神经系统中的作用与机制[J].神经解剖学杂志,2010, 26(5): 561—563.
[5]华海婴,李艳瑛.毛冬青甲素药理作用的研究[J].中国现代医药杂志,2006,8(11) :137—138.
[6]罗新海.毛冬青甲素治疗缺血性脑血管病. [J].广东医学, 1985, 64:26—28.
[7]江苏新医学院编.中医药大辞典.上册.[S]上海人民出版社, 1977: 441—443.
[8] Nacher J,Rosell DR,Alonso-Liosa G,MeEwen BS. NMDA receptor antagonist treatment induces a long-lasting increase in the number of proliferating cells PSA-NCAM-immunoreactive granule neurons and radial glia in the adult rat dentate gyrus.Eur J Neurosci.2001,13(3):512—520.
[9]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without cranicetomy in rats[J].Stroke,1989,20(1):84—91.
[10]方舒东,朱也森.脑缺血再灌注损伤的病理生理研究进展[J].医学综述,2006,12(18):1114—1116.
[11]韩敏,刘艳凯,薛茜.脑缺血再灌注损伤的研究进展[J].河北北方学院学报,2006,23(5):75—78.
[12]冯加纯,饶明俐,张淑琴,等.大鼠全脑缺血再灌注后几个脑区在不同时间的LPO、SOD、GSH-Px含量变化[J].中风与神经疾病杂志,1994,11(3):129—132.
[13]郭玉璞.缺血性脑损伤的病理生理基础和脑梗塞的治疗原则[J].基础医学与临床,1998,18(2):1—4.
[14]Lledo PM ,Alonso M , Grubb MS, et al . Adult neurogenesis and functional plasticity in neuronal circuits . Nat Rev Neurosci, 2006,7: 179—193.
[15]Alvarez-Buylla A, Garcia-Verdugo JM. Neurogenesis in adult subventricular zone. J Neurosci. 2002 ,22(3):629-634.
[16]Pencea V, Bingaman KD, Freedman LJ, Luskin MB. Neurogenesis in the subventricular zone and rostral migratory stream of the neonatal and adult primate forebrain. Exp Neurol. 2001,172(1):1-16.
[17]Kornack DR, Rakic P. The generation, migration, and differentiation of olfactory neurons in the adult primate brain.Proc Natl Acad Sci U S A. 2001,98(8):4752-4757.
[18]Sehara Y, Hayashi T, Deguchi K, et al . Distribution of inducible nitric oxide synthase and cell proliferation in rat brain after transient middle cerebral artery occlusion. Brain Res ,2006 , 1093 : 190—197.
[19]Li Y,Chopp M.Temporal profile of nestin expression after focal cerebral ischemia in adult rat[J].Brain Res,1999,838(1-2):1—10.
[20]Barres BA. New roles for glia[J]. J Neurosci, 1991, 11 (12) :3685—94.
[21]王娟,周义成,陈信坚.脑静脉窦血栓形成的MRI诊断[J].中国临床医学影像学杂志,2005,16(1):1008—1062.
[22] Loubeyre P,De Jaegere T,Tran-Minh VA.Three-dimensional phase contrast MR cerebral venography with zero filling interpolation in the slice encoding direction[J].Magn Reson Imaging,1999,17(8):1227—1233.
[23]Chiasson BJ,Tropepe V,Morshead CM ,van der Kooy D.Adult mammalian forebrain ependymal and subependymal cells demonstrate proliferative potentia1,but only subependymal cells have neura1 stem cell characteristics[J].J Neurosci,1999,19(11):4462—4471.
[24]Stevenson J,Knopp EA,Litt AW .MP-RAGE subtraction venography:a new technique[J].J Magn Reson Imaging,1995,5(2):239—241.
[25] Doetsch F.A niche for adult neural stem cells[J].Curr Opin Genet Dev,2003,13(5):543—550.
[26] Meister B,Bautz F,Brugger W,et a1.Expression of vascular endothelial growth factor(VEGF)and its receptors in human Neuroblastoma[J].Eur J Cancer,1999,35(3):445—449.
[27]焦淑洁,许慧芳,许杰,等. VEGF在人胚胎干细胞向神经元分化中作用的研究[J].Stroke and Nervous Diseases,2009,16(3):134—137.
[28]Hansen TM, Moss AJ, Brindle NP. Vascular endothelial growth factor and angiopoietins in neurovascular regeneration and protection following stroke[J]. Curr Neurovasc Res, 2008, 5(4): 236—245.
[29]Shen F, Fan Y, Su H, et al. Adeno-associated viral vector-mediated hypoxia-regulated VEGF gene transfer promotes angiogenesis following focal cerebral ischemia in mice[J]. Gene Ther, 2008,15(1): 30—39.
[30] Svensson B,Peters M,Poppe M,et a1.Vascular endothelial growth factor protects cultured rat hippocampal neurons against hypoxic injury via an antiexcitotoxic,caspase-independent mechanism[J].J Cereb Blood Flow Metab,2002,22(10):1170—1175.
[31]Sun Y,Jin K,Xie L,et a1.VEGF-induced neuroprotection,neurogenesis,and angiogenesis after focal cerebral ischemia[J].J Clin Invest,2003 Jun,111(12):1843—1851.
[32]杨树法等,血管内皮生长因子对胚胎干细胞来源神经干细胞增殖的促进[J].中国临床康复2006,10(1);12—14.
[33]彭好等,神经生长因子对新生鼠缺氧缺血性脑损伤神经修复作用的研究[J].中国新生儿科杂志,2007,22(2):85—88.
[34]董斌,罗其中,徐英辉.脑缺血半暗带区研究进展[J].中华神经外科疾病研究杂志,2004,4(1):90—92.
[35] Higashida RT,Furlan AJ,Roberts H,et al.Trial design and reporting standards for intra-arterial cerebral thrombolysis for acute ischemic stroke[J].Stroke,2003,34(8):109—137.
[36]陈洁文,谭宝璇,苏文.毛冬青甲素对脑细胞缺氧损伤的保护及其机制的实验研究[J].广州中医药大学学报,1997,14(2):97.
[37] Magavi SS, Macklis JD.Manipulation of neural precursorsin situ tower induction of neurogenesis in the adult brain:potential and limitation and limitation.Clin Neurosci Res,2002, 2 (1-2) : 40—57.
[38] Zhang PB, L iu Y, L i J , et al. Ependymal / subventricular zone cells migrate to the peri-infarct region and differentiate into neurons and astrocytes after focal cerebral ischemia in adult rats. Di Yi Jun YiDa Xue Xue Bao, 2005,25 (10) : 1201—1206.
[39] Yan JP, Liu XF, Liu H J, et al. Extracellular signal regulated k-inase involved in NGF /VEGF induced neuroprotective effect[J].Neurosci Lett, 2008, 434: 212—217.
[1] Tsyb AF, Roshal LM, Yuzhakov VV, et al. Morphofunctional study of the therapeutic effect of autologous mesenchymal stem cells in experimental diffuse brain injury in rats.Bull Exp Biol Med 2006;142(1):140—147.
[2] Wacharaprechanont T. Fetal stem cell: from research to clinical use.J Med Assoc Thai 2005;88 Suppl 2:S133—137.
[3]zhang YW, Denham J,Thies RS. Oligodendrocyte progenitor cells derived from human embryonic stem cells express neurotrophic factors.Stem Cells Dev 2006;15(6): 943—952.
[4] Li Y, Chopp M, Chen J et al. Intrastriatal transplantation of bone marrow nonhematopoietic cells improves functional recovery after stroke in adult mice. J Cereb Blood Flow Metab 2000;20(9):1311—1319.
[5] Sanchez-Ramos JR, Song S, Kamath SG,et al. Expression of neural markers in human umbilical cord blood. Exp Neurol 2001; 171(1):109—115.
[6] Chen J, Sanberg PR, Li Y, et al. Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats.Stroke 2001;32: 2682—2688.
[7]Yang L, Zheng J, Wang C. The in vitro study of the human adipose tissue-derived stromal cells differentiating into the neuron-like cells.Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2006;20(8):783—786.
[8] Rehman J, Traktuev D, Li J,ET al. Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation2004;109:1292—1298.
[9] Lathia JD,Rao MS,Mattson MP,et al. The microenvironment of the embryonic neural stem cell: Lessons from adult niches?Dev Dyn 2007;236(12):3267—3282.
[10] Hwang WS, Roh SI, Lee BC et al. Patient-specific embryonic stem cells derived from human SCNT blastocysts. Science 2005;308:1777—1783.
[11] Priddle H, Jones DK, Burridge PW, et al. Hematopoiesis from human embryonic stem cells: overcoming the immune barrier in stem cell therapies. Stem cells 2006;24(4):815—824.
[12] Bieberich E, Silva J, Wang G, et al. Selective apoptosis of pluripotent mouse andhuman stem cells by novel ceramide analogues prevents teratoma formation and enriches for neural precursors in ES cell-derived neural transplants.J Cell Biol 2004;167:723—734.
[13] Coyne TM, Marcus AJ, Woodbury D,et al. Marrow stromal cells transplanted to the adult brain are rejected by an inflammatory response and transfer donor labels to host neurons and glia. Stem Cells 2006;24(11):2483—2492.
[14] Ruth Kirschstein MD. Assessing Human Stem Cell Safety stem cells:scientific progress and future research ditections. Washington,DC, USA: Department of Health and Human Services 2001:93—99. [15 ]Chen J , Li Y, Chopp M. Intracerebral transplantation of bone marrow with BDNF after MCAO in rat. Neuropharmacology 2000; 39:711—716.
[16] Zhao LR, Duan WM , Reyes M , et al. Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp Neurol 2002;174(1):11—20. [17 ]Savitz SI, Rosenbaum DM , Dinsmore JH, et al. Cell transplantation for stroke . Ann Neurol 2002;52: 266—275.
[18] Chen J , Li Y, Wang L, et al. Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke 2001; 32(4): 1005—1011.
[19] Newman MB, Willing AE, Manresa JJ, et al. Cytokines produced by cultured human umbilical cord blood (HUCB) cells: implications for brain repair Exp Neurol 2006 ;199(1):201—208.
[20] Kondziolka D, Wechsler L, Goldstein S, et al. Transplantation of cultured human neuronal cell for patients with stroke.Neurology 2000;55: 505—569.
[21] Wiiling AE , Vendrme M , Mallery J ,et al. Mobilized peripheral blood cells administered intravenously produce functional recovery in stroke.Cell Transplant 2003;12:449—454.
[22] Lee ST, Park JE , Lee K, et al. Noninvasive method of immortalized neural stem-like cell transplantation in an experimental model of Huntington,s disease. J Neurosci Methods 2006;152(1-2):250—254.
[23] Jin K, Sun Y, Xie L , et al. Comparison of ischemia directed migration of neural precursor cells after intrastriatal , intraventricular , or intravenous transplantation in the rat. Neurobiol Dis 2005;18:366—374.
[24]Verfaillie CM,SchwartI R,Reyes M,et al.Unexpected potential of adult stell cell[J].Ann NY Acad Sci,2003,996:231—234.
[25] Shyu WC,Lin SZ,Ching MF,et al.Intracerbral peripheral blood stem cell(CD34+)implantation induces Neuroplasticity by enhancing betal 1 integrin-mediated angiogenesis in chronic stroke rats[J].Neurosci, 2006.26(13): 3444—3453. [26 ]Li Y.Chen J,Zhang CL,et al.Gliosis and brain remodeling after treatment of stroke in rats with marrow stromal cells. Glia.2005,49(3):407—417.
[27] Zhu D Y,Lau L,Liu SH,et al.Activation of camp-response-element–binding protein(CREB)after focal cerebral ischemia stimulates neurogenesis in the adult dentate gyrus.Proc Natl Acad Sci USA,2004,101:9453—9457.
[28] Xin H,Li Y,Chen X,et al.Bone marrow stromal cells induce BMP2/4 production in oxygen-glucose-deprived astrocytes,which promotes an astrocytic phenotype in adult subventricular progenitor cells[J].Neurosci Res,2006,83(8):1485—1493.
[29] Zhang C,Li Y,Chen J,et al.Bone marrow stromal cells upregulate expression of bone morphogenetic proteins 2 and 4 gap junction protein connexin-43 and synaptophysin after stroke in rats[J].Neuroscience,2006,141(2):687—695.
[30] Hess DC,Abe T,Hill WD,et al.Hematopoietic origin of microglial and perivascular cells in brain[J],Exp Neural, 2004;186(2):134—144.
[31] Chen J,Li Y,Katakowski M,et al.Intravenous bone marrow stromal cell therapy reduces apoptosis and promotes endogenous cell proliferation after stroke in female rat[J].Neurosci Res,2003,73:778—786.
[32]步星耀,黄志起,张永福.骨髓基质细胞的生物学特性及应用研究[J].实用诊断与治疗杂志,2004,18(1):35—38.
[33] Bang OY,Lee,IS,Lee PH,et a1.Autologous mesenchymal stem cell transp1antati。n in stroke patients [J]. Ann Neurol,2005,57(6):874—882.